Neural stem cells

Abstract

A homogenous, symmetrically dividing population of adherent neural stem cells is obtained from ES cells or foetal or adult brain isolates, using an activator of a signalling pathway downstream of a receptor of the EGF receptor family, optionally in combination with an activator of a signalling pathway downstream of an FGF receptor. The neural stem cell population is highly pure and retains the ability to differentiate into neurons after in excess of 100 passages.

The invention claimed is: 
     
       1. A composition, comprising:
 neural stem cells; 
 EGF; and 
 FGF-2, 
 
       wherein the neural stem cells are in a feeder-free, adherent monolayer culture and wherein at least 95% of said cells are symmetrically-dividing neural stem cells and wherein no more than 1% of the cells are positive for the expression of markers from mature astrocytes, neurons and oligodendrocytes. 
     
     
       2. The composition of  claim 1 , wherein at least 97% of said neural stem cells are symmetrically-dividing neural stem cells. 
     
     
       3. The composition of  claim 1 , wherein the neural stem cells are characterized in that they are positive for the expression of RC2, 3CB2, BLBP and Sox-2. 
     
     
       4. The composition of  claim 3 , wherein the neural stem cells are further characterized in that they are positive for the expression of at least one of (i) GLAST, (ii) Pax-6, (iii) the neural precursor markers nestin or vimentin, (iv) the LewisX antigen, (v) Musashi-1 and (vi) prominin. 
     
     
       5. The composition of  claim 4 , wherein the neural stem cells are further characterised in that they are negative for the expression of at least one of Oct4 and Nanog. 
     
     
       6. The composition of  claim 4 , wherein the neural stem cells are positive for the expression of Sox-2, and negative for the expression of Sox-1. 
     
     
       7. A mouse neural stem cell line, wherein said neural stem cells are maintained in the presence of EGF and FGF-2 in a feeder-free, adherent monolayer culture and wherein at least 95% of said cells are symmetrically-dividing neural stem cells and wherein no more than 1% of cells in the line are positive for expression of GFAP or βIII tubulin. 
     
     
       8. A human neural stem cell line, wherein said neural stem cells are maintained in the presence of EGF and FGF-2 in a feeder-free, adherent monolayer culture and wherein at least 95% of said cells are symmetrically-dividing neural stem cells and wherein no more than 1% of cells in the line are positive for expression of markers for mature astrocytes, neurons or oligodendrocytes. 
     
     
       9. A population of neural stem cells, wherein said neural stem cells are maintained in the presence of EGF and FGF-2 in a feeder-free, adherent monolayer culture and wherein at least 95% of said cells are symmetrically-dividing neural stem cells and wherein no more than 1% of the cells are positive for the expression markers for mature astrocytes, neurons, and oligodendrocytes. 
     
     
       10. A cell line, wherein the cells are maintained in the presence of EGF and FGF-2 in a feeder-free, adherent monolayer culture, wherein at least 95% of said cells are symmetrically-dividing neural stem cells and wherein no more than 1% of the cells are positive for the expression of markers for mature astrocytes, neurons, and oligodendrocytes. 
     
     
       11. A composition comprising neural stem cells, EGF, and FGF-2, wherein the neural stem cells are in a feeder-free, adherent monolayer culture and at least 95% of the cells in the composition are symmetrically-dividing neural stem cells and wherein no more than 1% of the cells are positive for the expression of markers for mature astrocytes, neurons, and oligodendrocytes. 
     
     
       12. A composition comprising neural stem cells, EGF, and FGF-2, wherein the neural stem cells are in a feeder-free, adherent monolayer culture, and wherein at least 95% of the cells in the composition are symmetrically-dividing neural stem cells.